Microorganisms programmed to produce immune modulators and anti-cancer therapeutics in tumor cells

Abstract

Genetically programmed microorganisms, such as bacteria or virus, pharmaceutical compositions thereof, and methods of modulating and treating cancers are disclosed.

The invention claimed is: 
     
       1. A genetically engineered non-pathogenic microorganism for intratumoral administration comprising a gene sequence for producing kynureninase, wherein the gene sequence is operably linked to an inducible promoter, and wherein the bacterium further comprises an aroP, tnaB, or mtr gene sequence encoding a transporter for importing kynurenine into the bacterium; and wherein the bacterium is an auxotroph comprising a deletion in a thyA, dapD, or dapA gene. 
     
     
       2. The genetically engineered bacterium of  claim 1 ,
 i) wherein the bacterium is a Gram-negative bacterium or a Gram-positive bacterium; 
 ii) wherein the bacterium is an obligate anaerobic bacterium, or a facultative anaerobic bacterium, or an aerobic bacterium; 
 iii) wherein the bacterium is a tumor-targeting bacterium; and/or 
 iv) wherein the bacterium is selected from  E. coli  Nissle,  Clostridium novyi  NT,  Clostridium butyricum , and  E. coli  K-12. 
 
     
     
       3. The genetically engineered bacterium of  claim 1 ,
 wherein the inducible promoter is induced by low-oxygen or anaerobic conditions, 
 wherein the inducible promoter is induced by the hypoxic environment of a tumor; 
 wherein the inducible promoter is a temperature sensitive promoter; and/or 
 wherein the inducible promoter is selected from a FNR-inducible promoter, an ANR-inducible promoter, a DNR-inducible promoter. 
 
     
     
       4. The genetically engineered bacterium of  claim 1 ,
 wherein the bacterium comprises a kill switch. 
 
     
     
       5. A pharmaceutically acceptable composition comprising the bacterium of  claim 1  and a pharmaceutically acceptable carrier, wherein the composition is formulated for intratumoral administration. 
     
     
       6. A method of treating or modulating cancer in a subject in need thereof comprising the step of administering to the subject the composition of  claim 1 . 
     
     
       7. The method of  claim 6 , further comprising administering to the subject a checkpoint inhibitor.